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新冠病毒核衣壳和受体结合域(RBD)混合抗原在植物中的生产及特性鉴定:一种针对COVID-19的候选疫苗

Production and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in Plant as a Vaccine Candidate against COVID-19.

作者信息

Mamedov Tarlan, Yuksel Damla, Ilgın Merve, Gürbüzaslan Irem, Gulec Burcu, Mammadova Gulshan, Ozdarendeli Aykut, Yetiskin Hazel, Kaplan Busra, Islam Pavel Shaikh Terkis, Uygut Muhammet Ali, Hasanova Gulnara

机构信息

Department of Agricultural Biotechnology, Akdeniz University, Antalya 07058, Turkey.

Department of Microbiology, Medical Faculty, Erciyes University, Kayseri 38280, Turkey.

出版信息

Vaccines (Basel). 2021 Nov 17;9(11):1337. doi: 10.3390/vaccines9111337.

Abstract

The COVID-19 pandemic has put global public health at high risk, rapidly spreading around the world. Although several COVID-19 vaccines are available for mass immunization, the world still urgently needs highly effective, reliable, cost-effective, and safe SARS-CoV-2 coronavirus vaccines, as well as antiviral and therapeutic drugs, to control the COVID-19 pandemic given the emerging variant strains of the virus. Recently, we successfully produced receptor-binding domain (RBD) variants in the plant as promising vaccine candidates against COVID-19 and demonstrated that mice immunized with these antigens elicited a high titer of RBD-specific antibodies with potent neutralizing activity against SARS-CoV-2. In this study, we engineered the nucleocapsid (N) protein and co-expressed it with RBD of SARS-CoV-2 in plant to produce an antigen cocktail. The purification yields were about 22 or 24 mg of pure protein/kg of plant biomass for N or N+RBD antigens, respectively. The purified plant produced N protein was recognized by N protein-specific monoclonal and polyclonal antibodies demonstrating specific reactivity of mAb to plant-produced N protein. In this study, for the first time, we report the co-expression of RBD with N protein to produce a cocktail antigen of SARS-CoV-2, which elicited high-titer antibodies with potent neutralizing activity against SARS-CoV-2. Thus, obtained data support that a plant-produced antigen cocktail, developed in this study, is a promising vaccine candidate against COVID-19.

摘要

新冠疫情使全球公共卫生面临高风险,在全球迅速蔓延。尽管有几种新冠疫苗可用于大规模免疫接种,但鉴于该病毒出现了变异毒株,世界仍迫切需要高效、可靠、具有成本效益且安全的严重急性呼吸综合征冠状病毒2(SARS-CoV-2)疫苗以及抗病毒和治疗药物来控制新冠疫情。最近,我们成功在植物中生产了受体结合域(RBD)变体,作为有前景的抗新冠疫苗候选物,并证明用这些抗原免疫的小鼠产生了高滴度的RBD特异性抗体,对SARS-CoV-2具有强大的中和活性。在本研究中,我们对核衣壳(N)蛋白进行了工程改造,并使其与SARS-CoV-2的RBD在植物中共表达,以产生一种抗原混合物。对于N或N+RBD抗原,纯化产量分别约为每千克植物生物量22毫克或24毫克纯蛋白。纯化后的植物产生的N蛋白可被N蛋白特异性单克隆和多克隆抗体识别,证明单克隆抗体对植物产生的N蛋白具有特异性反应性。在本研究中,我们首次报道了RBD与N蛋白的共表达,以产生SARS-CoV-2的混合抗原,该抗原引发了对SARS-CoV-2具有强大中和活性的高滴度抗体。因此,所获得的数据支持本研究中开发的植物产生的抗原混合物是一种有前景的抗新冠疫苗候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c57e/8621474/dda87be70f53/vaccines-09-01337-g001.jpg

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